Example Problems Example 1 Linear DNA DNA Sample Fragment Lengths bp Uncut DNA 10000 DNA cut with EcoRI 8000 2000 DNA cut with BamHI 5000 5000 DNA cut with EcoRI ID: 430466
Download Presentation The PPT/PDF document "Restriction Mapping" is the property of its rightful owner. Permission is granted to download and print the materials on this web site for personal, non-commercial use only, and to display it on your personal computer provided you do not modify the materials and that you retain all copyright notices contained in the materials. By downloading content from our website, you accept the terms of this agreement.
Slide1
Restriction Mapping
Example ProblemsSlide2
RFLP Analysis
Restriction Fragment Length Polymorphisms
Restriction enzymes from bacteria cut DNA molecules to create DNA fragments of varying lengths
Separate fragments in gel electrophoresisDNA fingerprinting (CSI, paternity)Slide3
Example #1: Linear DNA
DNA Sample
Fragment Lengths (
bp)Uncut DNA10,000DNA cut with EcoRI8,000 2,000DNA cut with BamHI5,000 5,000DNA cut with EcoRI + BamHI5,000 3,000 2,000Slide4
Example #1: Linear DNA
DNA Sample
Fragment Lengths (
bp)Uncut DNA10,000DNA cut with EcoRI8,000 2,000DNA cut with BamHI5,000 5,000DNA cut with EcoRI + BamHI5,000 3,000 2,000Uncut 10,000 bpSlide5
Example #1: Linear DNA
DNA Sample
Fragment Lengths (
bp)Uncut DNA10,000DNA cut with EcoRI8,000 2,000DNA cut with BamHI5,000 5,000DNA cut with EcoRI + BamHI5,000 3,000 2,000Uncut 10,000 bpEcoRI 8,000 bp2,000 bpSlide6
Example #1: Linear DNA
DNA Sample
Fragment Lengths (
bp)Uncut DNA10,000DNA cut with EcoRI8,000 2,000DNA cut with BamHI5,000 5,000DNA cut with EcoRI + BamHI5,000 3,000 2,000Uncut 10,000 bpEcoRI 8,000 bp2,000 bpBamHI
5,000 bp
5,000
bpSlide7
Example #1: Linear DNA
DNA Sample
Fragment Lengths (
bp)Uncut DNA10,000DNA cut with EcoRI8,000 2,000DNA cut with BamHI5,000 5,000DNA cut with EcoRI + BamHI5,000 3,000 2,000Uncut 10,000 bpEcoRI 8,000 bp2,000 bpBamHI
5,000 bp
5,000
bp
E + B
2,000
bp
5,000
bp
3,000
bp
B
ESlide8
Example #2: Plasmid (circular) DNA
DNA Sample
Fragment Length (
bp)Uncut DNA17,000DNA cut with SmaI17,000DNA cut with HaeIII17,000DNA cut with PstI10,000 7,000DNA cut with SmaI + HaeIII12,000 5,000DNA cut with SmaI + PstI10,000 6,000 1,000DNA cut with HaeIII + PstI7,000 6,000 4,000Slide9
Example #2: Plasmid (circular) DNA
DNA Sample
Fragment Length (
bp)Uncut DNA17,000DNA cut with SmaI17,000DNA cut with HaeIII17,000DNA cut with PstI10,000 7,000DNA cut with SmaI + HaeIII12,000 5,000DNA cut with SmaI + PstI10,000 6,000 1,000DNA cut with HaeIII + PstI7,000 6,000 4,000Uncut17,000Slide10
Example #2: Plasmid (circular) DNA
DNA Sample
Fragment Length (
bp)Uncut DNA17,000DNA cut with SmaI17,000DNA cut with HaeIII17,000DNA cut with PstI10,000 7,000DNA cut with SmaI + HaeIII12,000 5,000DNA cut with SmaI + PstI10,000 6,000 1,000DNA cut with HaeIII + PstI7,000 6,000 4,000Uncut17,000SmaI17,000
SSlide11
Example #2: Plasmid (circular) DNA
DNA Sample
Fragment Length (
bp)Uncut DNA17,000DNA cut with SmaI17,000DNA cut with HaeIII17,000DNA cut with PstI10,000 7,000DNA cut with SmaI + HaeIII12,000 5,000DNA cut with SmaI + PstI10,000 6,000 1,000DNA cut with HaeIII + PstI7,000 6,000 4,000Uncut17,000SmaI17,000
17,000
HaeIII
S
HSlide12
Example #2: Plasmid (circular) DNA
DNA Sample
Fragment Length (
bp)Uncut DNA17,000DNA cut with SmaI17,000DNA cut with HaeIII17,000DNA cut with PstI10,000 7,000DNA cut with SmaI + HaeIII12,000 5,000DNA cut with SmaI + PstI10,000 6,000 1,000DNA cut with HaeIII + PstI7,000 6,000 4,000Uncut17,000
PstI7,000
10,000
SmaI
17,000
17,000
HaeIII
S
H
P
PSlide13
Example #2: Plasmid (circular) DNA
DNA Sample
Fragment Length (
bp)Uncut DNA17,000DNA cut with SmaI17,000DNA cut with HaeIII17,000DNA cut with PstI10,000 7,000DNA cut with SmaI + HaeIII12,000 5,000DNA cut with SmaI + PstI10,000 6,000 1,000DNA cut with HaeIII + PstI7,000 6,000 4,000Uncut17,000SmaI17,000
17,000
HaeIII
PstI
7,000
10,000
S + H
5,000
12,000
S
H
P
P
H
SSlide14
Example #2: Plasmid (circular) DNA
DNA Sample
Fragment Length (
bp)Uncut DNA17,000DNA cut with SmaI17,000DNA cut with HaeIII17,000DNA cut with PstI10,000 7,000DNA cut with SmaI + HaeIII12,000 5,000DNA cut with SmaI + PstI10,000 6,000 1,000DNA cut with HaeIII + PstI7,000 6,000 4,000Uncut17,000PstI
7,00010,000
S + H
5,000
12,000
H
S
P
P
P
P
S
1,000
6,000
10,000
S + P
SmaI
17,000
17,000
HaeIII
S
HSlide15
Example #2: Plasmid (circular) DNA
DNA Sample
Fragment Length (
bp)Uncut DNA17,000DNA cut with SmaI17,000DNA cut with HaeIII17,000DNA cut with PstI10,000 7,000DNA cut with SmaI + HaeIII12,000 5,000DNA cut with SmaI + PstI10,000 6,000 1,000DNA cut with HaeIII + PstI7,000 6,000 4,000Uncut17,000PstI
7,00010,000
S + H
5,000
12,000
S
H
P
P
P
P
S
1,000
6,000
10,000
S + P
7,000
P
P
4,000
6,000
H
H + P
SmaI
17,000
17,000
HaeIII
S
HSlide16
Example #2: Plasmid (circular) DNA
DNA Sample
Fragment Length (
bp)Uncut DNA17,000DNA cut with SmaI17,000DNA cut with HaeIII17,000DNA cut with PstI10,000 7,000DNA cut with SmaI + HaeIII12,000 5,000DNA cut with SmaI + PstI10,000 6,000 1,000DNA cut with HaeIII + PstI7,000 6,000 4,000Uncut17,000PstI
7,00010,000
S + H
5,000
12,000
S
H
P
P
P
P
S
1,000
6,000
10,000
S + P
7,000
P
P
4,000
6,000
H
H + P
1,000
P
S
P
6,000
4,000
6,000
H
SmaI
17,000
17,000
HaeIII
S
H